The following chemicals were used: methanol, acetonitrile, isopropanol (HPLC grade, Merck, Darmstadt, Germany), formic acid (>96.0%), 2,2-azinobis-(ethyl-2,3-dihydrobenzothiazoline-6-sulphonic acid) diammonium salt-ABTS, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-Trolox, caffeic acid, ferulic acid, sinapic acid, quercetin, kaempferol, indole-3-carbinol, indole-3-acetonitrile, 3,3′-diindolylmethane, N-acetyl-l -cysteine, allyl isothiocyanate, sulforaphane (Sigma Aldrich, St. Louis, MO, USA), and glucotropaeolin (AppliChem, Darmstadt, Germany).
2 2 azino bis
Manufactured by Merck Group
Sourced in United States, Germany, Italy, Canada, Poland, India, Portugal, Spain, France, China, Australia, Sao Tome and Principe, Argentina
2,2'-azino-bis is a chromogenic substrate used in colorimetric assays. It is commonly used in enzyme-linked immunosorbent assays (ELISA) to detect and quantify the presence of specific analytes in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Diammonium salt, by Merck Group (65 mentions)
Gallic acid, by Merck Group (59 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (52 mentions)
Trolox, by Merck Group (47 mentions)
Quercetin, by Merck Group (42 mentions)
Potassium persulfate, by Merck Group (40 mentions)
Folin ciocalteu reagent, by Merck Group (32 mentions)
Methanol, by Merck Group (24 mentions)
Ascorbic acid, by Merck Group (23 mentions)
Formic acid, by Merck Group (19 mentions)
Hydrochloric acid (hcl), by Merck Group (19 mentions)
Ferulic acid, by Merck Group (18 mentions)
Sodium carbonate, by Merck Group (18 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (16 mentions)
Sodium hydroxide (naoh), by Merck Group (16 mentions)
Bovine serum albumin (bsa), by Merck Group (16 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (16 mentions)
Ethanol, by Merck Group (15 mentions)
Catechin, by Merck Group (15 mentions)
Acetonitrile, by Merck Group (15 mentions)
284 protocols using 2 2 azino bis
1
Phytochemical Screening and Antioxidant Evaluation
2
Antioxidant Evaluation of Phytochemicals
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The chemicals used in this work were all analytical grade. Ethylenediamine tetraacetic acid (EDTA) was purchased from Fluka (Steinheim, Germany),while copper sulfate pentahydrate (CuSO4.5 H2O), and ferrozine were acquired from Merck (Darmstadt, Germany).Butylated hydroxytoluene (BHT), quercetin, 1,1-diphenyl-2-picrylhydrazyl (DPPH), rutin hydrate, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, phosphoric acid, and pyrocatechol violet (PV). Phosphate buffered saline (PBS), trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), thiobarbituric acid (TBA), 2.2’-azobis (2-methylpropionamidine) dihydrochloride (AAPH), sulforhodamine B, and ellipticine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ethanol was purchased from Riedel de Haën (Buchs, Switzerland). Additional reagents and solvents were obtained from VWR International (Leuven, Belgium).
3
Enzymatic Assays for Oxidative Biocatalysis
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The activity of UPO was analyzed through the conversion of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (Merck KGaA, Darmstadt, Germany) to the ABTS radical cation (ε418 = 36,000 mM−1 cm−1), two molecules of 2,6-dimethoxyphenol (DMP) (Merck KGaA, Darmstadt, Germany) to coerulignone (ε469 = 49,600 mM−1 cm−1) and 5-nitro-1,3-benzodioxole (NBD) (Merck KGaA, Darmstadt, Darmstadt, Germany) to 4-nitrocatechol (ε425 = 9,700 mM−1 cm−1). For the activity assay, 5 μL–20 µL of sample were added to 85 μL–70 µL of assay buffer. The assay buffer compositions were as follows: ABTS-Assay : McIlvaine buffer pH 4.5, 0.3 mM ABTS, 10 mM GSSG; DMP-Assay : 0.1 M potassium phosphate buffer pH 6 (MroUPO) or pH 7 (AaeUPO), 6 mM DMP; NBD-Assay : McIlvaine buffer pH 7, 0.5 mM NBD. The reactions were started by adding 10 µL of 0.03% H2O2. Extinction measurements were performed using a microplate reader (Mithras2 LB 943 Multimode Plate Reade, Berthold Technologies GmbH & Co. KG).
4
Yeast Transformation and Enzyme Assays
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Yeast Transformation Kit, p-nitrophenyl butyrate and p-nitrophenyl β-d -galactopyranoside, High Pure Plasmid Isolation Kit, ABTS (2,2′azinobis (3ethylbenzothiazoline- 6 sulphonic acid)), p-methoxybenzyl alcohol, and Horseradish peroxidise (HRP) were purchased from Merck. Restriction enzymes NotI and BamHI were obtained from New England Biolabs. Phusion High-Fidelity DNA polymerase was obtained from NEB and QIAquick gel extraction kit from QIAGEN. Zymoprep™ Yeast Plasmid Miniprep II was purchased from Zymo Research. S. cerevisiae BJ5465 strain was purchased from LGC Promochem (Barcelona, Spain).
5
Antioxidant Activity Assays Protocol
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All the reagents used for the antioxidant activity assays, including Folin–Ciocâlteu phenol reagent, sodium carbonate (Na2CO3; 99% purity), tannic acid (Ph Eur purity), 1,1-diphenyl-2-picryl-hydrazyl (DPPH; 95% purity), 2,2′-azino-bis(3-thylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS; 98% purity), 2,2′-azobis (2-methylpropionamidine) dihydrochloride (AAPH; 97% purity), Trolox (97% purity), tert-butyl hydroperoxide solution (tBOOH; 80% purity) and the solvent ethanol (EtOH; 99.5% purity), were purchased from Merck (Darmstadt, Germany). To perform the assays, Trolox and DPPH were dissolved in EtOH, and ABTS and AAPH were prepared in phosphate-buffered saline (PBS; 0.1 M; pH = 7.0), sodium carbonate in sodium hydroxide (0.1 N) and tBOOH in water.
6
Antioxidant Potential of Plant Extracts
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All chemicals and reagents used in this study were of analytical or HPLC grade. 2,2-Diphenyl-1-picryhydrazyl hydrate stable radical (DPPH•, 95%), 2,2-azino-bis(3-ethyl- benzothiazoline-6-sulfonic acid) diammonium salt (ABTS, 98%), 2.0 M Folin–Ciocalteu phenol reagent, KCl, Na2HPO4, K2S2O8, NaCl, Na2CO3, and HPLC grade acetonitrile were purchased from Merck (Darmstadt, Germany); KH2PO4 from Jansen Chimica (Beerse, Belgium); 2,4,6-tripyridyl-s-triazine (TPTZ) from Fluka Chemicals (Steinheim, Switzerland). Acetone, methanol (MeOH), n-hexane and acetic acid were obtained from StanLab (Lublin, Poland); agricultural origin ethanol (96.6%) from Stumbras (Kaunas, Lithuania). Dimethyl sulfoxide (DMSO), n-butanol and ethyl acetate were purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Reference compounds, mangiferin and hydroxytormentic acid were purchased from Sigma Aldrich (Steinheim, Germany), and tormentic acid from ChemFaces (Wuhan, Hubei, China).
7
Antioxidant Compound Characterization
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Ethanol, mEthanol, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), potassium persulfate (K2S2O8), Lysogeny Broth with Agar and Thiazolyl Blue Tetrazolium Bromide (MTT) were from Merck (Darmstadt, Germany). Gentamicin sulfate was purchased from Biochrome.
8
Antimicrobial and Antioxidant Evaluation of Essential Oils
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EOs and HYs from inflorescences of L. nobilis, S. officinalis and S. sclarea cultivated in Tuscany (no wild site), Italy and obtained by steam distillation for 3 h (S. officinalis and S. sclarea) and 7 h (L. nobilis) extraction time, were directly provided by “èssenziale” Azienda Agricola, San Donato in Poggio (FI), Italy. Data of collection of plants: January 2020 for L. nobilis and June 2020 for S. officinalis and S. sclarea. All plLB Broth with Agar and Thiazolyl Blue Tetrazolium Bromide (MTT) were from Merck (Darmstadt, Germany). Gentamicin sulfate was purchased from Biochrom PAN-Bio-Tech GmbH (Aidenbach, Germany). Methanol, 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and potassium persulfate (K2S2O8) were from Merck.
9
Polyphenol Quantification and Antioxidant Assessment
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HPLC-grade acetonitrile (LC-MS grade), methanol, ethanol, petroleum ether, formic acid (>98% purity), Folin–Ciocalteu’s phenol reagent, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH•), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTSTM), potassium persulphate, (±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), gallic acid, quercetin, kaempferol, resveratrol, and rutin were supplied by Merck (Milan, Italy). De-ionized water (18.2 MΩ cm) was obtained from a Milli-Q purification system (Millipore, Bedford, MA, USA). quercetin 3-O-glucoside, kaempferol-3-O-rutinoside, kaempferol-3-O-glucoside, and hyperoside were supplied by Extrasynthese (Genay Cedex, France). Rhamnetin, isoRhamnetin, caftaric acid, and quercetin 3-O-glucuronide were from Phytolab (Vestenbergsgreuth, Germany).
10
Analytical Evaluation of Sweeteners
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All chemical reagents used in this study were of analytical grade. Ethanol 96 %, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), K2S2O8, Folin–Ciocalteu reagent, ascorbic acid, 3,5-dinitrosalicylic acid (DNS), sodium potassium tartrate, NaOH, Na2CO3, KCl, CH3COONa, KMnO4, 36.5 % HCl, glacial acetic acid, H2SO4 (0,1N) were purchased from Merck (Darmstadt, Germany). Ultrapure water was used in all the experiments. Sucrose was purchased from a local market in Cluj-Napoca, while all the other sweeteners used in this study were purchased from a health food store.
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